肌醇磷脂降解物在大鼠输精管平滑肌收缩中的作用

探讨肌醇磷脂代谢及其降解产物在大鼠输精管平滑肌收缩中的作用．用32P对输精管平滑肌进行标记，用薄层层析进行脂类分类，用放射自显影方法对磷脂进行定量研究，结果表明去甲肾上腺素能促进大鼠输精管平滑肌PI、PC、PA的代谢，PKC的大分子抑制剂和氯丙嗪不完全地抑制其代谢，Ca2+对上述反应有加速作用．本文提示去甲肾上腺素通过促进PI的代谢激活PKC，进而引起平滑肌的收缩，存在于输精管平滑肌中的PKC大分子蛋白抑制剂起着重要的调节作用．     

肝源性磷脂对小鼠学习记忆及海马窖蛋白-1表达的影响

以昆明种小鼠为研究对象,利用跳台实验、旷场实验和免疫印迹杂交等方法研究了肝源性磷脂对于增强小鼠学习记忆和空间探索能力的影响.结果表明:与对照组相比,预先给予高剂量肝磷脂补充可以显著提高小鼠的学习记忆以及空间探索能力;Western Blot结果显示,与正常组相比高剂量肝磷脂补充组小鼠脑内海马区Caveolin-1和PKCα蛋白的表达量明显升高.结果提示,肝源性磷脂可以通过激活Caveolin-1蛋白和PKCα蛋白的表达增强小鼠的学习和记忆能力.     

拟南芥PP2A B′调节亚基β亚型抗体的制备及鉴定

拟南芥中,蛋白磷酸酶2A(PP2A)B′调节亚基有9个亚型,其中α亚型和β亚型是油菜素内酯(BR)信号通路的正调节子,它们能使转录因子BZR1脱磷酸化.选择β亚型特异的一段多肽P109,利用大肠杆菌表达系统表达并纯化了连有HIS标签和GST标签的融合蛋白HIS-P109和GST-P109.以HIS-P109作为抗原,免疫新西兰兔,获得抗体,然后用GST-P109对抗体进行了亲和纯化.利用此纯化的抗体在不同的拟南芥材料中免疫印迹检测β亚型蛋白的表达,证实制备的抗体能与拟南芥PP2AB′调节亚基β亚型特异性反应,为深入研究PP2A在BR信号转导途径中的功能提供了有力工具.     

C-peptide stimulates Na<Superscript>+</Superscript>, K<Superscript>+</Superscript>-ATPase via activation of ERK1/2 MAP kinases in human renal tubular cells

Proinsulin-connecting peptide (C-peptide) exerts physiological effects partially via stimulation of Na⁺, K⁺-ATPase. We determined the molecular mechanism by which C-peptide stimulates Na⁺, K⁺-ATPase in primary human renal tubular cells (HRTCs). Incubation of the cells with 5 nM human C-peptide at 37°C for 10 min stimulated ⁸⁶Rb⁺ uptake by 40% (p<0.01). The carboxy-terminal pentapeptide was found to elicit 57% of the activity of the intact molecule. In parallel with ouabain-sensitive ⁸⁶Rb⁺ uptake, C-peptide increased subunit phosphorylation and basolateral membrane (BLM) abundance of the Na⁺, K⁺-ATPase ₁ and ₁ subunits. The increase in BLM abundance of the Na⁺, K⁺-ATPase ₁ and ₁ subunits was accompanied by depletion of ₁ and ₁ subunits from the endosomal compartments. C-peptide action on Na⁺, K⁺-ATPase was ERK1/2-dependent in HRTCs. C-peptide-stimulated Na⁺, K⁺-ATPase activation, phosphorylation of ₁-subunit and translocation of ₁ and ₁ subunits to the BLM were abolished by a MEK1/2 inhibitor (20 M PD98059). C-peptide stimulation of ⁸⁶Rb⁺ uptake was also abolished by preincubation of HRTCs with an inhibitor of PKC (1 M GF109203X). C-peptide stimulated phosphorylation of human Na⁺, K⁺-ATPase subunit on Thr-Pro amino acid motifs, which form specific ERK substrates. In conclusion, C-peptide stimulates sodium pump activity via ERK1/2-induced phosphorylation of Thr residues on the subunit of Na⁺, K⁺-ATPase.Received 15 June 2004; received after revision 14 September 2004; accepted 14 September 2004     

v-Fos transformation effector binds with CD2 cytoplasmic tail

The CD2 molecule, an adhesive and co-stimulating molecule, expresses virtually on all T lymphocytes, thymocytes, as well as natural killer cells, playing an important role in thymus development, T cell activation and apoptosis[1―3]. CD2 molecule consists of an ex- tracellular region with two Ig-like domains, a single membrane-spanning domain, and a positively charged proline-rich cytoplasmic tail, which is highly conserved between different species[1,4―7]. The functions of CD2 in signal t…     

An improved digital watermarking scheme combining with PKC

1　Introduction　WiththerapiddevelopmentoftheInternettechnique,electroniccommercebecomesmoreandmorepopular,whichbringsusmoreconvenience.Whileelectroniccommerceisproceeding,networkalsoprovidestheconvenientpassagefortheillegalcopies.Digitalwatermarkingistheenablingtechnologytoproveanownershiponcopyrightedproducts,detecttheoriginatorofillegallycopies,monitortheusageofthem,andanalyzethespreadspectrumofthedataovernetworksandservers.Fingerprintingisthetechniquewhichembedstheuniquebuyeridentificatio…     

Prokaryotic expression and characterization of a pea actin isoform (PEAc1) fused to GFP

~~Prokaryotic expression and characterization of a pea actin isoform (PEAc1) fused to GFP~~     

ERKs are the point of divergence of PKA and PKC activation by PTHrP in human skin fibroblasts

Parathyroid hormone-related peptide (PTHrP) receptors, coupled to trimeric G proteins, operate in most target cells through at least three different transduction routes: Gαs-mediated stimulation of adenylylcyclase (AC), Gαq-mediated activation of phospholipase Cβ (PLC) and mitogen-activated protein kinase (MAPK) activation. In this study we investigated the relative role of different pathways in human skin fibroblast prolifera-tion. Using chemical inhibitors and activators of signal transduction, we demonstrated that: (i) AC/cAMP and PLC/1,4,5 inositol triphosphate/diacylglycerol second-messenger systems are simultaneously activated following PTHrP binding to its receptors; (ii) the mitogenic response to PTHrP derives from a balance between two counteracting pathways – an activating route mediated by protein kinase C (PKC) and an inhibitory route mediated by protein kinase A (PKA); (iii) PTHrP mitogenic effects are largely dependent on MAPKs, whose activity can be modulate d by both PKA and PKC. Our results indicate that MAPKs are common targets of both transduction routes and, at the same time, their point of divergence in mediating PTHrP dual and opposite mitogenic effects. Received 2 August 2002; received after revision 10 September 2002; accepted 18 October 2002 RID="*" ID="*"Corresponding author.     

微丝解聚对DNA合成的影响

G_0期细胞在10％血清培养液刺激后向S期行进的早期出现微丝解聚的变化.细胞松弛素B(CB)0.5mg·L~_(-1)使微丝发生部分解聚能加强血清对DNA合成的刺激作用.但随着CB剂量的加大,微丝解聚的加强,便逐步增强对DNA合成的抑制作用.发现用5,10mg·L~(-1)的CB短时间处理G_0期细胞可强烈刺激蛋白激酶C(PKC)活性,虽使微丝暂时完全解聚,但不影响向S期的过渡及DNA的合成.促癌剂TPA(12-tetradecanoyl phorbol-13-acetate)可刺激G_0期细胞PKC活性,并能促进血清对DNA合成的刺激作用,但小剂量CB和TPA促进DNA的合成均依赖于血清的存在。     

植物谷氨酰胺合成酶研究进展

谷氨酰胺合成酶(GS)是参与高等植物氨同化过程的关键酶．介绍了高等植物谷氨酰胺合成酶及其同工酶的种类、性质、生理作用和分子生物学等方面的研究进展及在植物抗盐性方面的作用。